1-Cyclohexene-1-carboxaldehyde, 2-chloro-3-(hydroxymethylene)-, (3E)-

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(3E)-2-chloro-3-(hydroxymethylidene)cyclohex-1-ene-1-carbaldehyde

EC Number:

801-656-8

Cas Number:

1416808-92-8

Molecular formula:

C8H9ClO2

IUPAC Name:

(3E)-2-chloro-3-(hydroxymethylidene)cyclohex-1-ene-1-carbaldehyde

Test material form:

other: yellow solid molten

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

Dose range finding: up to 5000 µg/plate in the absence and presence of S9-mix in the strains TA100 and WP2uvrA
Main test: up to limited concentrations/plate because of observed toxicity in dose range finding.

Controls

Untreated negative controls:

yes

Remarks:

DMSO

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

2-nitrofluorene

sodium azide

methylmethanesulfonate

other: ICR-191 (Acros Organics, Geel, Belgium); 2-aminoanthracene (only control with all strains in presence of S9 mix)

Results and discussion

Test results

Test results 1

Species / strain:

bacteria, other: DOSE RANGE FINDING TEST: Salmonella typhimurium TA100 and Escherichia coli WP2uvrA

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

TA100: in the absence and presence of S9-mix

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

TA100: at 512 μg/plate and above in the absence of S9-mix and at 1600 and 5000 μg/plate in the presence of S9-mix; WP2uvrA: at 1600 and 5000 μg/plate in the absence and presence of S9-mix

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

bacteria, other: FIRST MAIN TEST: TA1535, TA1537 and TA98

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

TA98: in the presence of S9-mix

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

TA1535, TA1537 and TA98

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

bacteria, other: SECOND MAIN TEST: TA100, TA1535, TA1537, TA98 and WP2uvrA

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

TA100: in the absence and presence of S9-mix; TA98: in the presence of S9-mix

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

TA1535, TA1537 and TA98 in the absence of S9-mix and TA100 in the absence and presence of S9-mix

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

bacteria, other: THIRD MAIN TEST: TA1535, TA1537, TA98 and WP2uvrA

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

TA98: in the presence of S9-mix

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

all tester strains, except WP2uvrA in the presence of S9-mix

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

bacteria, other: FOURTH MAIN TEST: WP2uvrA

Metabolic activation:

with

Genotoxicity:

negative

Remarks:

in the presence of 10% (v/v) S9-mix and up to 5000 µg/plate

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

in the presence of 10% (v/v) S9-mix and up to 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Since 1.7- to 2.9-fold, dose related increases were observed in two tester strains, in the absence (TA100) and presence of S9-mix (TA98 and TA100) and the results were reproducible in the repeat experiments, these increases are biologically relevant and the substance is considered to be mutagenic in the absence and presence of S9-mix.

All other bacterial strains showed negative responses over the entire dose range, i.e. no dose-related, increase in the number of revertants in follow-up experiments.

In this study, the negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.

Applicant's summary and conclusion

Conclusions:

Interpretation of results : positive